Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1218, Issue 25, Pages 3845-3853Publisher
ELSEVIER
DOI: 10.1016/j.chroma.2011.04.044
Keywords
Magnetic polymer; Phosphate group; Enrichment of phosphopeptides; Mass spectrometry
Funding
- National Science Fund for Distinguished Young Scholars [20625516]
- Science Fund for Creative Research Groups [20921062]
- National 973 project of China [2007CB914200]
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As one of the most important post-translational modifications (PTM), reversible phosphorylation of protein is involved in many cellular processes. Enrichment and separation of phosphopeptides have become essential for large-scale identification of protein phosphorylation by mass spectrometry. In this work, five magnetic polymer materials with different numbers of phosphate groups were fabricated using a simple polymeric method and their abilities to enrich phosphopeptides were investigated. Our results showed that the enrichment efficiency is closely related to the number of phosphate groups attached to magnetic polymer sorbent. Under optimized condition (3% trifluoroacetic acid and 80% acetonitrile), magnetic polymer-particles with appropriate proportion of phosphate groups (Fe3O4@p(VPA-EDMA-1)-Zr4+) showed high performance for extracting phosphopeptides from complex peptides mixture of standard protein digestion. In this regard, a total of 988 unique phosphopeptides were successfully identified from proteolytic digestion of HeLa cell extracts by employing magnetic polymer-particles combined with nano-RPLC-MS/MS analysis. (C) 2011 Elsevier B.V. All rights reserved.
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